恶性黑素瘤细胞周期素D3及增殖细胞核抗原的表达

目的探讨恶性黑素瘤中细胞周期素D3和增殖细胞核抗原(PCNA)的表达及临床意义。方法采用免疫组化SP法检测57例原发性皮肤恶性黑素瘤及37例转移性恶性黑素瘤中细胞周期素D3和PCNA的表达，并以20例良性痣作为对照。结果①原发性皮肤恶性黑素瘤及转移性恶性黑素瘤中细胞周期素D3的阳性表达及PCNA的表达较良性痣均有不同程度增强：转移性恶性黑素瘤中两者的表达较原发性皮肤恶性黑素瘤也有不同程度增强。②两者在原发性皮肤恶性黑素瘤中的表达与病理Clark分级及淋巴结转移状态显著相关。③浅表扩散性恶性黑素瘤中细胞周期素D3阴性组的3年存活率显著高于细胞周期素D3阳性组，浅表扩散性恶性黑素瘤及结节性恶性黑素瘤中PCNA低表达组3年存活率明显高于PCNA高表达组。④浅表扩散性恶性黑素瘤及转移性恶性黑素瘤中细胞周期素D3阳性表达与PCNA高表达呈正相关。结论细胞周期素D3及PCNA的表达可能与原发性皮肤恶性黑素瘤的发生发展有关，对于治疗方案的选择及预后评估都具有重要的临床意义，细胞周期素D3的表达水平可作为浅表扩散性恶性黑素瘤临床预后的预测因子。     

胰岛素样生长因子ⅡmRNA结合蛋白3在恶性黑素瘤及良性痣中的表达

目的 探讨胰岛素样生长因子Ⅱ mRNA结合蛋白3(IMP3)在良性痣及黑素瘤组织中的表达,及其在恶性黑素瘤进展及诊断中的作用.方法 用IMP3抗体对28例恶性黑素瘤、8例Spitz痣、6例发育不良性痣和25例良性痣患者的标本组织进行免疫组化研究.结果 28例恶性黑素瘤组织标本中23例IMP3阳性,8例Spitz痣中4例阳性,6例发育不良性痣中2例阳性,25例良性痣均不表达.IMP3在黑素瘤中的表达明显高于Spitz痣及发育不良性痣(P<0.05),侵袭性黑素瘤表达明显高于原位黑素瘤(P<0.01).结论 IMP3可能是良性痣发展至恶性黑素瘤的一个生物学标志,在鉴别黑素瘤和良性痣之间存在一定的价值.     

恶性黑素瘤Survivin和C-erbB-2的检测及其意义

目的检测恶性黑素瘤的Survivin和CerbB2及其与临床病理特征的关系。方法应用免疫组化SP法检测72例恶性黑素瘤、61例瘤旁组织、22例黑色素细胞痣和20例正常皮肤组织中Survivin和CerbB2蛋白。结果恶性黑素瘤Survivin蛋白阳性率为59.7%,明显高于瘤旁组织3.3%,黑色素细胞痣4.5%和正常皮肤组织0%(P均<0.01)。Survivin阳性与转移状况和组织学分级有显著相关性。恶性黑素瘤无转移组和Ⅰ级分别明显低于转移组和Ⅱ,Ⅲ,Ⅳ级(P均<0.05)。恶性黑素瘤CerbB2蛋白阳性率为68.1%,明显高于瘤旁组织8.3%,黑色素细胞痣5.6%和正常皮肤组织2.8%(P均<0.01),CerbB2阳性与临床病理特征无明显相关性。Survivin在CerbB2阳性和阴性患者中阳性率分别为69.4%和39.1%,两者间有显著性差异(P<0.05)。结论Survivin和CerbB2蛋白在恶性黑素瘤组织中水平上调,提示两者对恶性黑素瘤发生发展起重要作用。恶性黑素瘤中Survivin蛋白水平与肿瘤转移和组织学分级有关,Survivin可能为临床判断恶性黑素瘤转移和评估预后的有用指标。     

骨桥蛋白在恶性黑素瘤中的表达

目的探讨骨桥蛋白在人恶性黑素瘤中的表达及意义,分析其与临床病理参数的关系。方法采用免疫组化SP法检测23例原发性恶性黑素瘤、17例转移性恶性黑素瘤及20例色素痣中骨桥蛋白的表达。结果骨桥蛋白在原发性恶性黑素瘤、转移性恶性黑素瘤、色素痣中的阳性表达率分别为91.3%,82.4%,15.0%,前两者阳性表达率明显高于后者,差异均有显著性意义(P均<0.05);骨桥蛋白的表达与年龄、性别、发病部位、是否淋巴结转移等病理因素均无关系(P均>0.05)。结论骨桥蛋白的表达可能与人恶性黑素瘤的侵袭行为的形成有关。     

乙酰肝素酶mRNA和蛋白在恶性黑素瘤皮损及A375细胞株中的表达

目的探讨乙酰肝素酶mRNA和蛋白在恶性黑素瘤皮损及A375细胞株中的表达及意义。方法选择恶性黑素瘤30例、黑素细胞痣30例、恶性黑素瘤A375细胞株及15例正常皮肤组织,分别采用原位杂交、免疫组化法检测皮损及细胞株中乙酰肝素酶mRNA及蛋白的表达。结果A375细胞株及19例(19/30,63.33%)恶性黑素瘤皮损中乙酰肝素酶mRNA呈阳性表达,2例(2/30,6.67%)黑素细胞痣皮损中乙酰肝素酶mRNA表达阳性,正常皮肤组织中无1例呈阳性表达;恶性黑素瘤皮损中乙酰肝素酶蛋白的阳性表达率与黑素细胞痣、与正常对照组相比,差异均有显著性意义(χ12=19.200,P=0.000;χ22=25.714,P=0.000)。乙酰肝素酶蛋白在恶性黑素瘤、恶性黑素瘤A375细胞株、黑素细胞痣及正常皮肤组织的表达情况与mRNA相一致。结论乙酰肝素酶mRNA和蛋白的高表达可能与恶性黑素瘤的发生发展有关,有望作为治疗黑素瘤的靶点。     

IGFBP7在皮肤黑素瘤组织和细胞系中的表达

目的:检测胰岛素样生长因子结合蛋白7(IGFBP7)在皮肤黑素瘤组织和细胞系中的表达。方法:免疫组化法检测25例原发性皮肤黑素瘤、5例转移性皮肤黑素瘤、20例良性色素痣、10例正常皮肤组织和4株人黑素瘤细胞系中IGFBP7的表达。结果:IGFBP7在正常皮肤和良性色素痣细胞浆中的阳性表达率为85%,而在皮肤黑素瘤胞浆中的阳性表达率为16%和20%。除人黑素瘤细胞系MV3阳性表达IGFBP7外,其余细胞系均阴性表达。结论:IGFBP7在皮肤黑素瘤组织和细胞系中的低表达或功能缺失可能参与了皮肤黑素瘤的发病。     

磷酸化活化转录因子2和磷酸化细胞信号传导与转录活化因子3在皮肤恶性黑素瘤中的表达

目的:探讨磷酸化活化转录因子(p-ATF)2和磷酸化细胞信号传导与转录活化因子(p-STAT)3在皮肤恶性黑素瘤中的表达及其临床意义.方法:采用免疫组化方法分别检测p-ATF2 和p-STAT3 在25 例原发性皮肤恶性黑素瘤(CMM)患者和14例转移性恶性黑素瘤(转移性MM)患者组织病理切片中的表达水平.结果:p-ATF2和p-STAT3在25例CMM和14例转移性MM 患者组织病理切片中细胞表达阳性率均分别显著高于色素痣中的表达阳性率(P ＜ 0.01),但在25 例CMM 和14 例转移性MM之间细胞表达阳性率差异无统计学意义(P ＞ 0.05).在25例CMM和14例转移性MM 患者组织病理切片中,p-ATF2阳性表达与p-STAT3阳性表达均分别有高度相关性(r分别为0.912和0.934,P 均＜0.01).结论:p-ATF2 和p-STAT3 虽在CMM 和转移性MM 中高表达,但与皮肤恶性黑素瘤的侵袭和转移无明显相关.     

人类单克隆抗体在固定组织切片上鉴别皮肤恶性黑素瘤和良性痣

本文作者用转移性黑素瘤患者局部淋巴结的淋巴细胞与小鼠骨髓瘤细胞株M5融合获得了6株稳定的杂交瘤,从中选出二株IgG单克隆抗体,采用直接卵蛋白—生物素—免疫过氧化酶染色方法研究了64个福尔马林固定,石腊包埋的组织切片.结果二株抗体均与所有18例原发性皮肤恶性黑素瘤、5例转移性皮肤黑素瘤及2例眼黑素瘤有阳性染色反应.其中一株抗体(2-139-1)能与4份恶性雀斑样痣标     

钠氢交换子调节因子-1及β-联蛋白在皮肤黑素瘤及痣细胞痣组织中的表达

目的 探讨皮肤黑素瘤及痣细胞痣组织中钠氢交换子调节因子-1 (NHERF1)和β-联蛋白表达的意义.方法 收集47例皮肤黑素瘤及37例痣细胞痣组织,用免疫组化方法对NHERF1和β-联蛋白在组织中的表达模式(膜表达、浆表达及核表达)和表达强度进行检测.结果 原位皮肤黑素瘤组织中NHERF1胞质中等/强阳性表达率为38％;而侵袭性皮肤黑素瘤组织中NHERF1胞质中等/强阳性表达率为71％,两组相比差异有统计学意义.同时,全部皮肤黑素瘤组织中NHERF1胞质中等/强阳性表达率为60％,而痣细胞痣组织中NHERF1胞质中等/强阳性表达率为0,差异有统计学意义.原位皮肤黑素瘤组织中可见β-联蛋白胞质中等/强阳性表达率为50％;而侵袭性皮肤黑素瘤组织中可见β-联蛋白胞质中等/强阳性表达率为84％.两组相比差异有统计学意义.全部皮肤黑素瘤组织中B一联蛋白胞质中等/强阳性表达率为72％,而痣细胞痣组织中NHERF1胞质中等/强阳性表达率为30％,两组相比具异有统计学意义.NHERF1及β-联蛋白胞质中等/强阳性表达率在痣细胞痣组、原位黑素瘤组及侵袭性黑素瘤组中均随着病变组织的恶性程度增加而升高.运用Spearman秩相关检验对皮肤黑素瘤中NHERF1及β-联蛋白的表达相关性进行分析显示无相关性.结论 NHERF1及β-联蛋白在痣细胞痣、原位黑素瘤及侵袭性黑素瘤组织中的胞质阳性表达随着病变组织的恶性程度增加而升高,但两者的表达差异无相关性.     

乙酰肝素酶、基质金属蛋白酶2和金属蛋白酶组织抑制剂2在恶性黑素瘤的表达

【摘要】 目的 探讨乙酰肝素酶、基质金属蛋白酶2（MMP2）和金属蛋白酶组织抑制剂2（TIMP2）蛋白在恶性黑素瘤皮损中的表达及意义。方法 选择恶性黑素瘤30例、黑素细胞痣30例、正常皮肤组织15例,分别采用免疫组化SP法检测皮损中乙酰肝素酶、MMP2和TIMP2蛋白的表达。结果 恶性黑素瘤皮损乙酰肝素酶蛋白的阳性表达率与黑素细胞痣、与正常皮肤组织相比,差异均有统计学意义（χ21 = 21.172,P < 0.01;χ22 = 27.805,P < 0.01）。恶性黑素瘤皮损MMP2蛋白的阳性表达率与黑素细胞痣、与正常皮肤组织相比,差异均有统计学意义（χ21 = 19.817,P < 0.01;χ22 = 19.866,P < 0.01）。恶性黑素瘤皮损TIMP2蛋白的阳性表达率与黑素细胞痣、与正常皮肤组织相比,差异均有统计学意义（χ21 = 19.200,P < 0.01;χ22 = 15.000,P < 0.01）。结论 乙酰肝素酶、MMP2和TIMP2蛋白在恶性黑素瘤中的表达水平明显高于其在黑素细胞痣和正常皮肤中的表达水平。     

Staining of melanocytic neoplasms by melanoma antigen recognized by T cells

We stained benign melanocytic nevi and malignant melanoma with antibodies to melanoma antigen recognized by T cells (Mart-1) to determine if this was useful in differentiating benign from malignant melanocytic neoplasms. Forty-five primary malignant melanomas and 71 benign melanocytic nevi were stained with antibodies to Mart-1. Two cases of malignant melanoma metastatic to lymph node and three cutaneous metastases of malignant melanoma were also stained. The degree of staining was graded into diffuse positive staining, focal positive staining, and negative staining. Thirty-six of 45 primary malignant melanomas stained diffusely positive with antibodies to Mart-1. This included three of five desmoplastic malignant melanomas that showed positive staining. Four melanomas showed faint or focal positive staining. One of two metastases to lymph node showed strong positive staining and one showed no staining. All three cutaneous metastases showed diffuse positive staining. Sixty-one of 71 melanocytic nevi showed no staining or faint staining with antibodies to Mart-1. Ten of 71 melanocytic nevi showed strong positive staining. The majority of these were congenital nevi. Staining with antibodies to Mart-1 antigen was a useful marker of malignant melanoma. However, staining may also be seen in benign melanocytic neoplasms. The presence or absence of staining for Mart-1 antigen cannot be used to differentiate benign melanocytic nevi from malignant melanocytic tumors.     

Proliferation antigens in cutaneous melanocytic tumors--an immunohistochemical study comparing the transferrin receptor and the Ki 67 antigen

The cellular reactivities with the monoclonal antibodies OKT9 and Ki 67 have been demonstrated to be closely related to proliferation in various malignant neoplasms. In this study a total of 25 melanocytic skin tumors was examined immunohistochemically with both antibodies and the results were evaluated semiquantitatively for OKT9 and quantitatively for Ki 67 by stereological methods. All cases of primary and metastatic malignant melanoma expressed a strong stainability for OKT9, whereas benign melanocytic nevi were almost completely negative. Our results with the monoclonal antibody Ki 67 revealed highly significant differences in the numerical density of Ki-67-positive cells between metastatic malignant melanoma (number of positive cells: 47.0 +/- 9.2 X 10(3)/mm3), primary malignant melanoma (6.3 +/- 1.9 X 10(3)/mm3) and benign melanocytic nevi (2.2 +/- 0.7 X 10(3)/mm3). Correlation analysis between mean percentage of OKT9-positive cells and numerical density of Ki-67-positive cells revealed a significant correlation of both parameters (r = 0.58; p less than or equal to 0.05), indicating a positive relationship of OKT9 and Ki 67 expression. Especially in primary malignant melanoma, however, the amount of OKT9-positive cells considerably exceeds that of Ki-67-positive cells. The monoclonal antibodies OKT9 and Ki 67 reflect 'proliferative activity' in melanocytic skin tumors, as both are expressed in significantly higher amounts in primary and metastatic malignant melanomas. The combined application of these antibodies in cutaneous melanocytic lesions might be of diagnostic and prognostic value.     

载脂蛋白D在皮肤恶性黑素瘤中的水平检测及临床意义

目的　检测皮肤恶性黑色素瘤(CMM)中载脂蛋白D(apoD)的水平并研究其临床意义。方法　用免疫组化SP法检测65例原发性CMM和20例黑色素细胞痣中apoD的水平。结果　CMM中apoD阳性25例(38. 46% ),所有黑色素细胞痣均阴性。apoD的阳性与患者的年龄、性别及肿瘤发生的部位无明显关系, 而与CMM的组织病理分型和Clark分级有关(P<0. 05)。apoD阳性患者,术后3年生存率显著低于阴性患者(P<0. 05)。结论　apoD参与了CMM的发生发展过程,可能是CMM预后不良的一个新的预测指标。     

Overexpression of the cyclin-dependent kinase inhibitor p21WAF1/GIP1 in human cutaneous malignant melanoma

p2l^WAFI/CIPI (p21) is an inhibitor of cyclin-dependent kinases recently identified as the downstream effector of wild-type p53-me-diated cell cycle arrest. The gene coding for p21 may function as a negative regulator of melanoma growth, progression, and metastasis. Using immunohistochemistry and Western blotting, we investigated the expression of p21 in human melanocytic proliferations. Immunohistochemical staining was performed on 13 common acquired nevi, 12 dysplastic nevi, 23 primary malignant melanomas, and 12 metastatic melanomas. Common acquired nevi showed minimal p21 staining (1.8±0.3%, mean±SEM). The percentage of positive nuclei was slightly elevated in dysplastic nevi (8.9±1.7%). Both primary malignant melanoma (29±3%) and metastatic melanoma (33±5%) demonstrated a significantly increased number of p21-positive nuclei compared to benign lesions (p<0.001). p21 was strongly expressed even in actively proliferating lesions as confirmed by MIB-1 labelling, and although the majority of p21-positive cells likely represent a non-proliferating population, staining was occasionally observed in cells undergoing mitosis, suggesting abnormal function of this cell cycle inhibitor in malignant melanoma. Overexpression of p21 in metastatic melanoma compared to common acquired nevi was confirmed by Western blot analysis of human tumor samples. These findings suggest that increased p21 expression relative to benign nevi is not sufficient to control melanoma growth in vivo.     

Malignant blue nevus with lymph node metastases

Background:  Malignant blue nevi arise within cellular blue nevi and contain atypical mitoses, necrosis, nuclear pleomorphism and prominent nucleoli. Malignant blue nevus has been described as a distinct identity, a rare form of malignant melanoma, and a misdiagnosed melanoma.
Methods:  We present a patient with metastatic malignant blue nevus and studies on the histopathologic, immunohistochemical, and molecular features of the neoplasm.
Results:  Histology showed a malignant blue nevus arising in a combined intradermal and cellular blue nevus. CD117 (c-kit) staining showed diffuse cytoplasmic expression within the cellular blue nevus, decreased staining in the malignant component, and variable positivity within the lymph node metastases. Molecular loss of heterozygosity analysis showed different allelic patterns at the hOGG-1 locus between the melanoma and control skin specimens with a varying heterozygous allelic pattern in both the benign and malignant blue nevus.
Conclusions:  Our case of malignant blue nevus with lymph node metastasis involved mutation of the hOGG-1 DNA repair gene. CD117 showed decreased staining of the primary malignant blue nevus with marked upregulation in the metastatic lesion, unlike most metastatic melanomas. Further study is needed to determine if hOGG-1 mutation or c-kit upregulation play a role in the pathogenesis of dendritic melanocytic lesions (either benign or malignant).     

黑素瘤细胞中脂肪酸结合蛋白7的表达及其与细胞增殖、凋亡的关系

目的探讨黑素瘤细胞中脂肪酸结合蛋白7(FABP7)的表达及其与细胞增殖、凋亡的关系。方法采用免疫组织化学方法分析临床45例良性痣(为皮内痣)、60例原位黑素瘤、60例转移性黑素瘤标本中FABP7的表达。取组织细胞进行体外培养,选择FABP7特异性小干扰RNA(siRNA)降低FABP7的表达,然后检测黑素瘤细胞的增殖及凋亡情况。结果良性痣细胞浆及细胞核中FABP7的表达均高于原位黑素瘤、转移性黑素瘤(P<0.05),原位黑素瘤和转移性黑素瘤细胞浆及细胞核中的FABP7表达差异无显著性(P>0.05)。下调FABP7表达可减少原位黑素瘤细胞31.0%及转移性黑素瘤细胞81.0%的增殖性,下调FABP7表达对原位黑素瘤及转移性黑素瘤细胞的凋亡率无明显影响。结论良性痣中FABP7表达比黑素瘤高,FABP7对体外培养黑素瘤细胞增殖有影响。     

Antiapoptotic bcl-2 and bcl-xL in advanced malignant melanoma

Apoptosis is an important cofactor in the pathogenesis of a plethora of malignancies. However, little is known about modulation of the expression of bcl gene family in melanocytic tumors. To determine the role of bcl-2, bcl-x and bax in melanocytic tumors we investigated the differential expression of these genes via RT-PCR in tissue samples from human benign nevi, primary melanomas and melanoma metastases in comparison with normal skin. Bcl-2 was strongly expressed in 14/16 metastases (87.5%), whereas only 7/13 primary melanomas (53%), 7/15 nevi (46%) and 7/16 normal tissue samples (43%) showed expression of bcl-2 (P < 0.05). There was a strong indication of a correlation between tumor thickness and bcl-2 expression in nodular malignant melanomas. Expression of bcl-x was found in 16/16 melanoma metastases (100%), 11/13 primary melanomas (84%), 12/15 nevi (80%) and 10/16 normal tissue samples (62%) (P < 0.05). Bcl-xL expression increased from primary melanoma to melanoma metastases, whereas bcl-xS showed a decreasing expression level during melanoma progression. No differences in bax expression were seen between melanoma metastases, primary melanoma, nevi and normal tissue. Immunohistochemical investigations of another 53 tissue samples showed similar results. Our results strongly indicate that bcl-2 and bcl-xL gene expression increases with progression of malignant melanoma. Bcl-2 and bcl-xL expression could reflect an increased malignant potential caused by an inhibition of apoptosis and growth advantage for metastatic melanoma cells.     

Expression analysis of ovostatin 2 reveals its involvement in proliferation,invasion and angiogenesis of cutaneous malignant melanoma

The relationship of ovostatin 2 (OVOS2) expression with the clinicopathological features of cutaneous malignant melanoma (CMM) was investigated to identify OVOS2 expression in cutaneous melanocytic lesions, and to reveal whether OVOS2 has a function in melanoma progression. Eight specimens of CMM and paracancerous tissue were analyzed using real‐time polymerase chain reaction (PCR) and western blot for the mRNA and protein expression of OVOS2, respectively. Immunohistochemical staining was performed on 52 CMM and 62 nevi, followed by clinicopathological significance analysis. The proliferative cells were visualized by staining with Ki‐67 antibody. The intensity of angiogenesis was assessed by staining with vascular endothelial growth factor (VEGF). Real‐time PCR and western blot analyses showed that OVOS2 was significantly upregulated in cutaneous melanoma than in paired normal skins. Immunohistochemistry showed that 86.5% (45/52) of malignant cases showed OVOS2 cytoplasmic expression compared with 29% (18/62) in benign nevi. OVOS2 expression was significantly higher in invasive and metastatic melanoma than in in situ melanoma (P < 0.01). Furthermore, OVOS2 expression was positively correlated with the known prognostic variables of melanoma including clinical stage, Clark level and Breslow depth. It was also significantly associated with ulcer status, Ki‐67 labeling index and VEGF expression in primary melanoma. OVOS2 expression was significantly increased in CMM, which increased incrementally from benign nevi to melanoma and appeared to be involved in the progression of melanoma.